title: Type 2M von Willebrand disease: F606I and I662F mutations in the glycoprotein Ib binding domain selectively impair ristocetin- but not botrocetin-mediated binding of von Willebrand factor to platelets.
abstract: von Willebrand disease (vWD) is a common, autosomally inherited, bleeding disorder caused by quantitative and/or qualitative deficiency of von Willebrand factor (vWF). We describe two families with a variant form of vWD where affected members of both families have borderline or low vWF antigen levels, normal vWF multimer patterns, disproportionately low ristocetin cofactor activity, and significant bleeding symptoms. Whereas ristocetin-induced binding of plasma vWF from affected members of both families to fixed platelets was reduced, botrocetin-induced platelet binding was normal. The sequencing of genomic DNA identified unique missense mutations in each family in the vWF exon 28. In Family A, a missense mutation at nucleotide 4105T --> A resulted in a Phe606Ile amino acid substitution (F606I) and in Family B, a missense mutation at nucleotide 4273A --> T resulted in an Ile662Phe amino acid substitution (I662F). Both mutations are within the large disulfide loop between Cys509 and Cys695 in the A1 domain that mediates vWF interaction with platelet glycoprotein Ib. Expression of recombinant vWF containing either F606I or I662F mutations resulted in mutant recombinant vWF with decreased ristocetin-induced platelet binding, but normal multimer structure, botrocetin-induced platelet binding, collagen binding, and binding to the conformation-sensitive monoclonal antibody, AvW-3. Both mutations are phenotypically distinct from the previously reported variant type 2MMilwaukee-1 because of the presence of normal botrocetin-induced platelet binding, collagen binding, and AvW-3 binding, as well as the greater frequency and intensity of clinical bleeding. When the reported type 2M mutations are mapped on the predicted three-dimensional structure of the A1 loop of vWF, the mutations cluster in one region that is distinct from the region in which the type 2B mutations cluster.
authors.affiliation: Blood Research Institute, The Blood Center of Southeastern Wisconsin, Milwaukee, WI 53233, USA.
authors.forename: C A
authors.initials: CA
authors.lastname: Hillery
authors.forename: D J
authors.initials: DJ
authors.lastname: Mancuso
authors.forename: J
authors.initials: J
authors.lastname: Evan Sadler
authors.forename: J W
authors.initials: JW
authors.lastname: Ponder
authors.forename: M A
authors.initials: MA
authors.lastname: Jozwiak
authors.forename: P A
authors.initials: PA
authors.lastname: Christopherson
authors.forename: J
authors.initials: J
authors.lastname: Cox Gill
authors.forename: J
authors.initials: J
authors.lastname: Paul Scott
authors.forename: R R
authors.initials: RR
authors.lastname: Montgomery
mesh_terms: D001483:Base Sequence; D001665:Binding Sites; D001792:Blood Platelets / Q000378:metabolism*; D003094:Collagen / Q000378:metabolism; D003435:Crotalid Venoms / Q000494:pharmacology*; D004247:DNA / Q000737:chemistry; D005260:Female; D006388:Hemagglutinins / Q000494:pharmacology; D006801:Humans; D008297:Male; D008958:Models, Molecular; D009154:Mutation; D010375:Pedigree; D019038:Platelet Glycoprotein GPIb-IX Complex / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012310:Ristocetin / Q000494:pharmacology*; D014842:von Willebrand Diseases / Q000235:genetics*; D014841:von Willebrand Factor / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
chemical_list: D003435:Crotalid Venoms; D006388:Hemagglutinins; D019038:Platelet Glycoprotein GPIb-IX Complex; D011994:Recombinant Proteins; D014841:von Willebrand Factor; D012310:Ristocetin; C030961:botrocetin; D003094:Collagen; D004247:DNA